Aim:To isolate, identify, and differentiate Capripoxviruses (CaPV) (sheep pox virus and goat pox virus) infections by egg inoculation, transmission electron microscopy (TEM), and 30 kDa RNA polymerase subunit gene-based polymerase chain reaction (PCR) (RPO30) in clinically affected animals in Hawamdia township of Giza Governorate, Egypt.Materials and Methods:A total of 37 scab samples were collected from clinically suspected field cases of sheep pox and goat pox. These samples were collected during (2014-2015) during different outbreaks of sheep pox and goat pox from Hawamdia township of Giza Governorate, Egypt. The samples were subjected to egg inoculation, TEM, and (RPO30) gene-based PCR. By using the egg inoculation: Previously prepared 37 scab samples (n=23 sheep and n=14 goats) were inoculated on the chorioallantoic membrane of specific pathogen free (SPF) embryonated chicken eggs (12 days old age). In the presence of the suitable percentage of humidity and candling, the inoculated eggs were incubated at 37°C. By using the TEM: Samples showed positive pock lesions on the chorioallantoic membranes, were fixed in glutaraldehyde, then processed and sectioned for TEM. Using the (RPO30) gene-based PCR assay, 30 of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened.Results:Using the egg inoculation, a characteristic pock lesions for poxviruses were seen in 30/37 (n=19 sheep and n=11 goats) (81.08%). Using the TEM, examination of the positive samples after egg inoculation revealed positive result in 23/30 (n=15 sheep and n=8 goats) (76.66%). The positive results represented by the presence of negatively stained oval-shape virus particles. Using the (RPO30) gene-based PCR assay, out of 30 total of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened, 27 (90%) samples (n=17 sheep and n=10 goats) were positive. The given band sizes of sheep and goats were 172 and 152 bp, respectively.Conclusion:PCR assay depended on RPO30 gene can be used lonely for the detection, identification, and differentiation of CaPVs. RPO30 gene-based PCR assay in combination with gene sequencing helps in molecular epidemiological studies of CaPV infection.
The present study was designed to investigate the chemoprotective effect of green tea extract (GTE), rosmarinic acid (RA) and rosemary extract (RE) against diethylnitrosamine (DEN) initiated and ferric nitrilotriacetate (Fe-NTA) promoted nephrotoxicity in rats. Forty male rats were categorized into five: Group I included healthy rats, group II received DEN+Fe-NTA, group III received 200 mg/kg b.wt. of RE+DEN+Fe-NTA, group IV received 1 g/kg b.wt. of GTE+DEN+Fe-NTA and group V received 50 mg/kg b.wt. of RA+DEN+Fe-NTA. RE, GTE, RA were given orally for 14 days before single intraperitoneal administration of DEN (160 mg/kg) till the end of the experiment. Eighteen days after DEN, a single intraperitoneal dose of Fe-NTA (5 mg Fe/kg) was administrated to rats to promote nephrotoxicity. The biochemical parameters were analyzed in serum at time intervals while the malondialdehyde (MDA) and tumor necrosis factor-alpha (TNF-α) were assessed in both serum and renal tissues. Kidney from each group was histopathologically examined at time intervals. The administration of Fe-NTA after DEN dose to albino rats resulted in acute nephrotoxicity which was characterized by a highly significant elevation of serum urea, creatinine, uric acid (p=0.000), serum and renal MDA and TNF-α (p=0.000) with vacuolation of epithelial lining renal tubules. The administration of RE, GTE and RA prior to DEN+Fe-NTA treatment significantly ameliorated the observed increased levels of the above mentioned parameters. GTE, RA & RE exerted a protective effect against renal toxicity with GTE showing a more pronounced effect on renal function parameters while RA showed the best antioxidant impact.
SUMMARY:The present study aims to produce low cost sophorolipids, and to evaluate their potential hypocholesterolemic impact. Sophorolipids were produced by Candida bombicola grown on safflower oil cake, extracted by methanol followed by ethyl acetate with a yield of 24.4 and 48.3 g·100 g −1 mixed substrate, respectively. Their structure was confirmed by FTIR and 1 H NMR and proven to be safe when subjected to an acute toxicity test. A biological experiment was done on 42 male albino rats classified into six groups for 4 weeks following an induction period for hypercholesterolemia of 8 weeks. The two extracts and their mixture were examined for their hypocholesterolemic effect compared to rosuvastatin. The results revealed a reduction in total cholesterol, low density lipoprotein cholesterol, atherogenic index, liver transaminases' activity and malondialdehyde. They also revealed an elevation in high density lipoprotein cholesterol and antioxidant enzymes which was more efficient than rosuvastatin. Histopathological examination confirmed these results. In conclusion, the newly isolated sophorolipids are powerful hypocholesterolemic compounds which are even more efficient and safer than rosuvastatin.KEYWORDS: Candida bombicola; Hypocholesterolemic; Lipid profile; Rats; Safflower oil cake; Solid state fermentation; Sophorolipids RESUMEN: Efecto hipocolesterolémico de soforolípidos recién aislados producidos por la conversión microbiana de la torta de aceite de cártamo en ratas alimentadas con una dieta rica en grasas y colesterol. El presente estudio tiene como objetivo producir soforolípidos de bajo costo, evaluando su potencial impacto hipocolesterolémico. Los soforolípidos fueron producidos por Candida bombicola cultivada en torta de aceite de cártamo, extraída con metanol seguido de acetato de etilo con un rendimiento de 24,4 y 48,3 g·100 g −1 de sustrato mixto, respectivamente. Su estructura fue confirmada por FTIR y 1 H RMN y demostró ser segura cuando se sometió a prueba de toxicidad aguda. Un experimento biológico se realizó con 42 ratones albinos machos clasificados en seis grupos, durante 4 semanas, después de un período de inducción al hipercolesterolemia de 8 semanas. Se examinaron los dos extractos y su mezcla para determinar su efecto hipocolesterolémico en comparación con rosuvastatina. Los resultados revelaron una reducción en el colesterol total, el colesterol de lipoproteínas de baja densidad, el índice aterogénico, la actividad de las transaminasas hepáticas y el malondialdehído, mientras que mostraron una elevación del colesterol de lipoproteínas de alta densidad y de las enzimas antioxidantes más eficientemente que la rosuvastatina. El examen histopatológico confirmó estos resultados. En conclusión, los soforolípidos recién aislados son potentes compuestos hipocolesterolémicos aún más eficientes y más seguros que la rosuvastatina.
Background and Aim: The peste des petits ruminants (PPR) is a highly contagious disease of small ruminants which negatively affects animal production and the socioeconomic status of farmers. Peste des petits ruminants virus (PPRV) encodes eight proteins, with the viral fusion protein (F) playing a role in virus virulence and stimulating an effective protective immune response. This study aimed to isolate and complete the identification of PPRV circulating in goats in different Egyptian governorates and perform molecular characterization of the PPRV F gene. Materials and Methods: Samples were collected from unvaccinated animals with clinical signs suggestive of PPR. A total of 256 sera were tested for the detection of PPRV antibodies using a competitive enzyme-linked immunosorbent assay (c-ELISA) kit, while 214 samples of blood buffy coat preparation, animal swabs (nasal, ocular, and saliva), and fecal and tissue samples were tested for the detection of the PPRV antigen using an antigen-capture ELISA kit. Molecular diagnosis, gene cloning, blast analysis, and phylogenetic analysis were performed for the molecular characterization of PPRV. Results: The seroprevalence results of PPRV antibodies in the tested sera showed a total of 67.9% positive samples. The rates of PPR antigen recorded by the antigen-capture ELISA in the swabs (nasal and ocular) and tissue samples were 44.3%, 46.8%, and 43.5%, respectively, with saliva swabs having the highest rate of PPRV positivity (76.4%) and fecal samples having the lowest (33.3%). Molecular characterization of the PPRV Vero cell culture revealed that the circulating PPRV strain belongs to the IV lineage. Blast analysis of the PPRV F gene showed 96.7% identity with the PPRV strain Egypt-2014 fusion protein (F) gene, KT006589.1, differing by 43 single-nucleotide polymorphisms. Conclusion: The results of this study indicate that the emerging PPRV belongs to the IV lineage among small ruminant animals. The findings also indicate the need for an innovative strategy to control and eliminate this disease based on a regularly administered and effective vaccine, a test to distinguish between infected and vaccinated animals, and the need for further study on the protein structure and PPRV F gene expression, which should help us to understand the molecular evolution of the virus and control and eliminate PPR disease.
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